Validating Microarray Data Using RT2 Real-TimeTM PCR Products

Real-time PCR monitors the amount of amplicon as the reaction occurs. Usually, the amount of product is directly related to the fluorescence of a reporter dye. Because it detects the amount of product as the reaction progresses, real-time PCR provides a wide linear dynamic range, demonstrates high sensitivity, and is very quantitative. The initial amount of template DNA is inversely proportional to a parameter measured for each reaction, the threshold cycle (Ct). While not requiring post-reaction processing (such as characterization by agarose gel electrophoresis), real-time PCR does require dedicated and expensive equipment. The method is expensive to set up initially, but it becomes rather cost-effective once used routinely particularly when employing SYBR Green as the fluorescent reporter.